The principal long-range goal is the development of methods for the total synthesis of biologically functional genes and to understand molecular mechanisms of gene control and expression. The total research involved in this program can be categorized as follows: (1) Further development of the chemical methods for the synthesis of deoxyribopolynucleotides. (2) Total synthesis of the structural gene corresponding to the entire length (126 nucleotides) of the precursor for the E. coli tyrosine tRNA. (3) Mechanism of the initiation of transcription by RNA polymerase using the tyrosine tRNA gene. (4) Total synthesis of the DNA region corresponding to the promoter region in tyrosine tRNA gene. (5) Mechanism of termination and the length of the terminator region in tyrosine tRNA gene. (6) Total synthesis of the DNA corresponding to the terminator region of the tyrosine tRNA gene. (7) Total synthesis of a functional tRNA gene possessing the transcriptional control elements. (8) Maturation of the primary transcript of the synthetic tyrosine tRNA gene to a functional tRNA in vitro. (9) Structure-function relationships in the tyrosine tRNA by structural modifications at the gene level. (10) In vivo studies of the totally synthetic bacterial tRNA gene. Studies will be performed to demonstrate suppression of the amber mutation in an E. coli mutant by the synthetic gene. (11) Nature and function of the bacterial membrane-DNA associations. BIBLIOGRAPHIC REFERENCES: Studies on Polynucleotides. CXXXII. Total Synthesis of the Structural Gene for the Precursor of a Tyrosine Suppressor Transfer RNA from E. coli (2). Chemical Synthesis of the Deoxypolynucloetide Segments Corresponding to the Nucleotide Sequence 1-31. Hans van de Sande, Marvin H. Caruthers, Ashok Kumar and H. Gobind Khorana, J. Biol. Chem., 251, 571 (1976). Studies on Polynucleotides. CXXXVI. Total Synthesis of the Structural Gene for the Precursor of a Tyrosine Suppressor Transfer RNA from E. coli (6). Synthesis of the Segments Corresponding to the Nucleotide Sequence 100-126. Kan L. Agarwal, Marvin H. Caruthers, Henry Buchi, J. Hans van de Sande and H. Gobind Khorana, J. Biol. Chem., 251, 624 (1976).